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e-ISSN: 2278-3008, p-ISSN:2319-7676. Volume 10, Issue 1 Ver. III (Jan -Feb. 2015), PP 20-26
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Polymorphism in Glutatione S-Transferase P1 and ManganeseSuperoxide Dismmutase Genes in Egyptian Women with
Preeclampsia
Hesham M. Alaa3, Amal K. Seleem1, Dalia Shaalan1, Abd Elaziz El-Refaeey2,
Hend Shalaby2 And Ayman S. Al-Hussaini3
Medical Biochemistry Department1, Obstetrics and Gynecology Department 2 Faculty of Medicine , Mansoura
University and Chemistry Department3, Faculty of Sience, Port Said University.
Abstract: Susceptibility to preeclampsia is believed to have a genetic component. Several studies have reported
associations between polymorphisms of oxidative stress- related genes and preeclampsia. The aim of the present
study was to study the polymorphisms in anti-oxidant genes glutathione S-transferase P1 (GSTP1) and Mnsuperoxide dismutase (Mn-SOD) in patients with of preeclampsia. Seventy four preeclampsia patients and fifty
age-matched healthy pregnant female controls were genotyped for GSTP1 and Mn-SOD. DNA was extracted
from peripheral blood of all patients and control women. DNA analysis was carried out by polymerase chain
reaction (PCR), and then digestion of the PCR products by restriction enzymes (RFLP) for both genes was
performed. As regards GSTP1, the carriers of Val (G) allele were significantly more frequent among
preeclampsia patients when compared to the control group (45.95% Vs. 19%, X2=10.40, OR=2.418, 95%CI
(1.3698 - 4.269); p=0.0023). Preeclampsia patients had a lower frequency of GSTP1 Ile/Ile (AA) genotype
(31.1% Vs. 68% in control; X2=13.09, OR=0.457, 95%CI (0.241 - 0.866); p =0.0164). The frequency of
GSTP1 Ile/Val (AG) and Val/Val (GG) genotypes was higher in preeclampsia patients than control (68.9%Vs.
32%; X2= 12.83, OR=2.154, 95%CI (1.106 - 4.194); p =0.0241). However, non significant frequencies
differences as regards Mn-SOD genotypes or alleles were found between preeclampsia patients and control. It
could be concluded that, pregnant Egyptian women carrying the Val (G) allele of GSTP1 GSTP1 -105 Ile →Val
(-313 A to G) polymorphism may be more susceptible to preeclampsia either in homozygous or heterozygous
state.
Keywords: Preeclampsia, GSTP1, Mn-SOD, gene polymorphism.
I.
Introduction
Preeclampsia is a potentially life-threatening disease that occurs exclusively in pregnant women during
late gestation (>20 weeks). The two hallmark symptoms are hypertension and proteinuria, complicating at least
5% of pregnancies and are usually resolved upon delivery of the placenta (Lain and Roberts, 2002). Although
preeclampsia is a leading cause of maternal death and a major contributor to maternal and prenatal morbidity, its
precise cause has not been completely elucidated (Chamy et al., 2006). Susceptibility to preeclampsia is
believed to have a genetic component and several studies have reported associations between polymorphisms of
oxidative stress and hypertension-related genes with preeclampsia. The mechanism of oxidative stressassociated hypertension has been continuously evolving with the popular theme of endothelial dysfunction. The
etiology of oxidative stress in preeclamptic women has therefore been the focus of extensive research (Zhang et
al., 2008). Low levels of reactive oxygen species (ROS) are crucial in maintaining normal cellular physiologic
functions, such as proliferation, apoptosis, and cell cycle arrest; whereas increased levels of ROS induce
oxidative stress and cause an imbalanced hemostatic microenvironment (Aggarwal and Gehlot, 2009).
The products of the glutathione S-transferase-P1 (GSTP1) and manganese superoxide dismutase ( MnSOD; SOD-2) genes are involved in the regulation of ROS metabolic processes and some single nucleotide
polymorphisms (SNPs) within these genes result in dysregulated protein translation and/ or altered function (Xu
et al., 2012). The GSTP1, a member of the glutathione S-transferase family of phase 2 detoxification isozymes,
is a candidate enzyme protecting epithelial cells from ROS-induced oxidative stress and detoxifying free
radicals by catalyzing the nucleophilic addition of hydrophobic and electrophilic compounds to reduced
glutathione (Zusterzeel et al., 1999). The GSTP1 gene, located at 11q13, has a single nucleotide polymorphic
site at codon 105 (exon 5). This SNP (rs1695) is a substitution of adenosine-to-guanosine at nucleotide 313 (A313-G) results in an isoleucine (Ile, ATC)-to-valine (Val, GTC) change at residue 105 (Ile-105-Val) (Zusterzeel
et al., 2000). This amino acid substitution, in the GSTP-1 binding site, decreases its catalytic activity and causes
less effective detoxification. The GSTP1 enzymes with Val-105 have altered heat stability and represented with
specific activity of the Val-containing isoform (Hurl et al., 2005).
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Polymorphism In Glutatione S-Transferase P1 And Manganese-Superoxide Dismmutase…
The Mn-SOD, SOD2, is one of the major superoxide scavengers in mitochondria, and it catalyzes
accumulated superoxide radicals into hydrogen peroxide (H2O2). The Mn-SOD gene contains five exons and
spans almost 20 kb located at 6p25. The 47 cytosine-to-thymine (C-47-T) transition at codon 16 in the Mn-SOD
gene (rs4880) creates a sense mutation of alanine (Ala, GCT)-to-valine (Val, GTT) (Ala-16-Val) in the 24amino acid signal sequence of Mn-SOD protein, which affects protein folding and localization (Cai et al., 2004;
Wang et al., 2009). This SNP has been identified in exon 2 of the human Mn-SOD gene. Molecularly, the Val
variant is predicted to form a β-sheet structure on the protein, while the Ala variant results in an α-helical
conformation. MnSOD is synthesized in the cytoplasm and transported into the mitochondria via an N-terminal
mitochondrial targeting sequence (MTS) (Zelko et al., 2002). The Ala variant is more efficiently imported into
the mitochondria than the Val variant. Miscoding of Ala to Val will result in arresting of Mn-SOD in the inner
mitochondrial membrane but not in the mitochondrial matrix, therefore reducing its functional activities (Sutton
et al., 2005). The inefficient Mn-SOD protein in the mitochondria may leave the cell vulnerable to oxidative
damage without its full defense against superoxide radicals, which leads to protein oxidation and DNA
mutations (Bag and Bag, 2008). Several studies have revealed that this Mn-SOD polymorphism is associated
with cardiovascular and metabolic diseases in the general population (Tian et al., 2011, Kariz et al., 2012).
Results of studies seeking associations of the Mn-SOD C-47-T and GSTP1 A-313-G polymorphisms
with preeclampsia have not always been consistent among different population analyses. Thus, our objective
was to clarify a possible association between these two SNPs and the risk for developing preeclampsia among
Egyptian women.
II.
Subject And Methods
Patient selection:
During the period from July 2011 to November 2012, a case- control study was conducted in Obstetric
and Gynecology and Medical Biochemistry Departments, Mansoura Faculty of Medicine. Seventy four
primigravida with preeclampsia with singleton pregnancy, were recruited from Obstetrics Unit of Obstetrics and
Gynecology Department. The control group included 50 age-matched healthy pregnant women with singleton
pregnancy free from any diseases. All participants underwent history taking, physical examination for
measurement of blood pressure, body weight, obstetric examination and obstetric ultrasonography. Basic
laboratory tests as complete blood count, serum creatinine, ALT, AST, and repeated urine analysis for
proteinuria were done. Informed consent was taken from each participant. The study was approved by The
Ethical Committee of Mansoura Faculty of Medicine, Egypt.
Preeclampsia was diagnosed when hypertension was accompanied with proteinuria (>300 mg/dl/24h)
Hypertension was defined when systolic blood pressure ≥140 mmHg and/or diastolic blood pressure ≥90 mmHg
measured on two consecutive occasions at least 6 hour apart.
Pregnant women with chronic hypertension, superimposed preeclampsia, patients with eclampsia were
excluded. Also, patients with gestational diabetes mellitus, associated diabetes mellitus, and chronic renal
disease, and twin pregnancies were excluded.
Biochemical investigation:
Venous blood samples were taken from all participants. Three ml were delivered to centrifuge tubes
containing K2EDTA (stored as EDTA anti-coagulated blood sample at -30ºC for DNA extraction). Another 5 ml
blood sample was allowed to clot for 15 minutes and centrifuged at 7000 rpm for 10 minutes for serum
separation to determine: serum creatinine, ALT and AST. Samples were centrifuged at 4000 xg for 20 min at
4ºC to avoid the contamination of the serum with platelets. They were then stored at –20ºC until used.
DNA extraction:
Genomic DNA was extracted from EDTA–anticoagulated peripheral blood leucocytes using QIA amp
DNA Blood Mini Kit supplied by Qiagen GmbH (Cat, No.51104, Hiden, Germany) (Schur et al., 2001). The
average DNA concentration (0.161±0.002µg/µl) was determined from absorbance at 260 nm (Jenway, Genova
Model, UK). All samples had a 260/280 nm absorbance ratio between 1.4 and 1.63. The integrity of the DNA
was checked by electrophoresis on 2 % agarose gel stained with ethidium bromide.
Genotyping of GSTP1 -105 Ile → Val (-313 A to G) SNP (rs1695) and Mn-SOD -16 Ala → Val (-47 C to T)
SNP (rs4880):
Conventional method of PCR amplification was used for genotyping of GSTP1 -105 Ile →Val (-313 A
to G) SNP (rs1695) in exon 5 of GSTP1 gene by the method described by Li et al. (2010). A 440 bp exon 5
region containing the polymorphism site was amplified by using the following primers sequences: a 20-mer
GSTP1- forward primer 5′- ACG CAC ATC CTC TTC CCC TC -3′ and a 20-mer reverse primer 5′- TAC TTG
GCT GGT TGA TGT CC -3′ (Fermentas Life Science, Ontario, Canada) in which the underlined nucleotide
DOI: 10.9790/3008-10132026
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Polymorphism In Glutatione S-Transferase P1 And Manganese-Superoxide Dismmutase…
represents the deliberated primer mismatch designed to introduce an artificial Bsm AI restriction site when the
G allele is present at position -313. Genotyping of Mn-SOD -16 Ala → Val (-47 C to T) SNP (rs4880) was
done by the method described by Manica-Cattani et al. (2012). A 110 bp exon 2 region containing the
polymorphism site was amplified by using the following primers sequences: a 22-mer forward primer 5′- ACC
AGC AGG CAG CTG GCG CCG G-3′ and a 20-mer reverse primer 5′-GCG TTG ATG TGA GGT TCC AG-3′
(Fermentas Life Science, Ontario, Canada) in which the underlined nucleotide represents the deliberated primer
mismatch designed to introduce an artificial Hae III restriction site when the C allele is present at position -47.
PCR was carried out in 50 microliters final reaction volume using Ready Mix (RED. Taq-PCR
Reaction Mix) (purchased from Sigma Aldrich, Saint Louis, USA).The following mixture was prepared for each
sample: 25 µl RED-Taq PCR reaction Mix (1X), 2µl (20 pmol) of forward primer, 2µl (20 pmol) of reverse
primer, 2µl (200 ng) of genomic DNA and 19 µl of deionized water. This mix was put in a thin wall PCR
microcentrifuge tube and gently centrifuged to collect all components to the bottom of the tube. Thermal cycling
was carried out using Techne TC-312 thermal cycler (Model FTC312D, Barlworld Scientific Ltd, UK). The
following program was used: initial denaturation at 94 ºC for 5 min followed by 35 cycles of denaturation at 94
ºC for 30 sec, annealing at 57 ºC (GSTP-1) or 60 ºC (Mn-SOD) for 30 sec, and an extension step at 72 ºC for 30
sec, followed by a final linear extension step at 72 ºC for 5 min. Amplification products of 440 bp for GSTP-1
and 110 bp for Mn-SOD were obtained when electrophoresed in a 2.5% (wt/vol) agarose gel stained with
ethidium bromide, and bands were visualized under ultraviolet (UV) via Light UV Transilluminator (Model
TUV-20, OWI Scientific,Inc.800 242-5560).
The PCR products of GSTP-1 (440 bp) were digested using Bsm AI enzyme (Fermentas Life Science,
Conventional restriction enzymes, Catalog number ER0031), 12μl of each PCR product was digested overnight
with 3 μl (30 U) of the enzyme at 37 °C. The enzyme recognition site is 5'-GTCTC (N)1-3' and 3'CAGAG(N)5-5'. Homozygous wild genotype (Ile/Ile; AA) was presented as a single band at 440 bp when
electrophoresed in a 2.5% (wt/vol) agarose gel against DNA molecular marker. Two bands at 228 bp and 212
bp identified Val/Val (GG) homozygous genotype, and three bands at 440, 228 and 212 bp indicated
heterozygous Ile/Val (AG) genotype (Figure 1).
To determine the Val (T) allele of Mn-SOD at position -47, PCR products of 110 bp were digested by
Hae III enzyme (Fermentas Life Science, Conventional restriction enzymes, Catalog number ER0151), 12μl of
each PCR product was digested overnight with 3 μl (30 U) of the enzyme at 37 °C. The enzyme recognition site
is 5'-GGCC-3' and 3'-CCGG-5'. A single band (110 bp) indicated the Val/Val (TT) genotype, while two bands at
85 bp and 25 bp identified the wild Ala/Ala (CC) genotype. The presence of all the three bands (110 bp, 85 bp
and 25 bp) indicated the Ala/Val (CT) genotype (Figure 2).
Statistical analysis:
Statistical package of social science (SPSS) version 16 was used for statistical analysis. The qualitative data
were presented in the form of number and percentage. Chi-square test was used as a test of significance for
qualitative data. Odds ratio and 95% confidence intervals were calculated for determination of disease
association and expected risk for the disease. The quantitative data were expressed as mean and standard
deviation. Significance was considered at p value less than 0.05.
III.
Results
Clinical and demographic data of 74 patients and 50 control are summarized in table (1). The mean age
of preeclampsia patients was 27.58±4.55 years with a mean age of gestation 29.47±2.58 weeks. Significant
differences were found in the frequency of GSTP1 -105 Ile →Val (-313 A to G) SNP genotypes among
preeclampsia patients and control as shown in table (2). Patients with preeclampsia had a lower frequency of
GSTP1 -105 Ile/Ile (AA) genotype (31.1% Vs 68% in controls ; OR=0.457 with 95%CI (0.241 - 0.866); p
=0.0164). The frequency of GSTP1 -105 Ile/Val (AG) genotypes was higher in preeclamptic patients than
control (45.9% Vs 26%; OR= 1.767 with 95% CI (0.849 - 3.678); p=0.0251). GSTP1 -105 homozygous
Val/Val (GG) genotype was found in 23% of preeclampsia patients and 6% of control carried that genotype. It
was significantly more frequent in preeclampsia patients than in control (OR=3.829 with 95% CI (1.066 13.754); p =0.0396).
The frequency of GSTP1 -105 allele's distribution among the preeclampsia patients and control are
shown in table (2). As regards carriers of Val (G) allele, it was significantly more frequent among preeclampsia
patients with 2.418 folds higher risk of developing preeclampsia among patients group than controls (OR=2.418,
95% CI=1.3698 - 4.269, p=0.0023). On the other hand, the carriers of Ile (A) allele were significantly more
frequent among control when compared to preeclampsia group (OR=0.667, 95% CI= 0.447 - 0.995; p=0.0474).
Table (3) shows that the relative risk for preeclampsia was 4.712 folds higher in patients carrying Ile →Val ( A
to G) transition either in homozygote or heterozygote form than those carrying the dominant homozygote Ile/Ile
(AA) form (OR=4.712, 95% CI=2.178-10.193, p=0.0001). Also, the relative risk for preeclampsia was higher in
DOI: 10.9790/3008-10132026
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Polymorphism In Glutatione S-Transferase P1 And Manganese-Superoxide Dismmutase…
patients carrying the Val (G) allele than those carrying the Ile (A) allele (OR=3.624, 95% CI= 1.999-6.570; p
<0.0001).
As regards Mn-SOD -16 Ala → Val SNP (-47 C to T) genotypes, no significant differences were found
in the genotype frequencies between patients and controls. Also, the frequency of Mn-SOD -16 Ala → Val
allele's distribution among the preeclampsia patients and control shows non significant differences (Table 4).
Neither of the Ala → Val transition genotypes nor alleles show any significant differences when compared to
the dominant ones as regards preeclampsia relative risk (Table 5).
Table (1) Clinical and demographic data of patients and control:
Variable
Age (years)
Gravity
Parity
Gestational age (weeks)
Preeclampsia (n=74)
27.58±4.55
2.45±1.24
1.24±1.18
29.47±2.58
Control
(n=50)
25.78±4.21
2.82±1.32
1.63±1.20
29.33±2.54
P value
0.052
0.1
0.06
0.6
Data are presented as mean ± SD
Table (2) Distribution of GSTP1 -105 Ile → Val (-313 A to G) SNP genotypes and Ile and Val (A and G)
alleles among patients and control:
GSTP1- 105 Ile → Val
genotypes
Control
(n=50)
No (%)
34 (68%)
X2
OR
(95%CI)
P value
Ile/Ile (AA)
Preeclampsia
(n=74)
No
(%)
23 (31.1%)
13.09
0.0164
Ile/Val (AG)
34 (45.9%)
13 (26%)
5.01
Val/Val (GG)
17 (23%)
3 (6%)
8.83
Ile/Val (AG)
+Val/Val (GG)
Ile (A)
51 (68.9%)
16 (32%)
12.83
80 (54.05%)
81 (81%)
5.01
Val (G)
68 (45.95%)
19 (19%)
10.40
0.457
(0.241 - 0.866)
1.767
(0.849 - 3.678)
3.829
(1.066 -13.754)
2.154
(1.106 - 4.194)
0.667
(0.447 - 0.995)
2.418
(1.3698 - 4.269)
0.0251
0.0396
0.0241
0.0474
0.0023
p is significant if < 0.05
Table ( 3 ): Association of GSTP1 -105 Ile → Val (-313 A to G) SNP genotypes and alleles to the relative
preeclampsia risk:
GSTP1 -105 Ile → Val
SNP
Ile/Ile (AA)
Preeclampsia
(n=74)
No
(%)
23 (31.1%)
Control
(n=50 )
No (%)
34 (68%)
Ile/Val (AG)
+Val/Val (GG)
Ile (A)
51 (68.9%)
16 (32%)
80 (54.05%)
81 (81%)
Val (G)
68 (45.95%)
19 (19%)
X2
OR
(95% CI)
P value
16.3
4.712
(2.178-10.193)
0.0001
19.02
3.624
(1.999-6.570)
0.0001
P is significant if < 0.05
Figure 1: Agarose gel electrophoretic analysis of the GSTP1 -105 Ile → Val (-313 A to G) (rs1695):
DOI: 10.9790/3008-10132026
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Polymorphism In Glutatione S-Transferase P1 And Manganese-Superoxide Dismmutase…
The genotypes of GSTP1 -105 Ile →Val (-313 A to G) SNP (rs1695) after Bsm AI digestion analysis
represented as follow: Ile/Ile (AA) genotype is presented by one single band at 440 bp. ( lane 1) , Ile/Val (AG)
genotype is presented by three bands at 440, 228 and 212 bp (lane 2) and.Val/Val (GG) genotype is presented by
two bands at 228 bp and 212 bp (lane 3) Lane (M) represents the molecular marker (DNA molecular weight
marker, purchased from Promega Technical Service, Catalog#G3161).
Table (4) Distribution of Mn-SOD -16 Ala → Val (-47 C to T) SNP genotypes and Ala and Val alleles (C
and T) among patients and control:
Mn-SOD -16 Val → Ala
genotypes
Control
(n=50)
No (%)
39 (78%)
X2
OR
(95%CI)
P value
Ala/Ala (CC)
Preeclampsia
(n=74)
No
(%)
53 (71.6%)
0.24
0.7600
Ala / Val (CT)
18 (24.3%)
10 (20%)
0.21
Val/Val (TT)
3 (4%)
1 (1%)
0.80
Ala / Val (CT)
+ Val/Val (TT)
Ala (C)
21 (28.4%)
11 (22%)
0.50
124 (83.78%)
88 (88%)
0.05
Val (T)
24 (16.22%)
12 (12%)
0.32
0.918
(0.531 - 1.588)
1.216
(0.519 - 2.852)
2.027
(0.205 - 20.046)
1.290
(0.572 - 2.908)
0.952
(0.656 - 1.382)
1.351
(0.646 - 2.827)
0.6526
0.5456
0.5394
0.7963
0.4239
p is significant if < 0.05
Table (5): Association of Mn-SOD -16 Ala → Val (-47 C to T) SNP genotypes and alleles to preeclampsia
risk:
Mn-SOD -16 Val → Ala
SNP
Ala/Ala (CC)
Ala / Val (CT)
+ Val/Val (TT)
Ala (C)
Val (T)
Preeclampsia (n=74)
No
(%)
53 (71.6%)
21 (28.4%)
124 (83.78%)
24 (16.22%)
Control
(n=50 )
No (%)
39 (78%)
11 (22%)
88 (88%)
12 (12%)
X2
OR
(95% CI)
P value
0.63
1.405
(0.607-3.250)
0.4269
0.85
1.420
(0.674-2.990)
0.3568
p is significant if < 0.05
Figure 2: Agarose gel electrophoretic analysis of the Mn-SOD -16 Ala → Val (-47 C to T) SNP (rs4880):
The genotypes of Mn-SOD -16 Ala → Val (-47 C to T) SNP (rs4880) after Hae III digestion analysis
represented as follow: Ala/Ala (CC) wild genotype is presented by two bands at 85 bp and 25 bp (lane 1),
Ala/Val (CT) genotype is presented by three bands at 110 bp, 85 bp and 25 bp (lane 2), and Val/Val (TT) mutant
genotype is presented by one single band at 110 bp ( lane 3). Lane (M) represents the molecular marker (DNA
molecular weight marker, purchased from Promega Technical Service, Catalog#G3161).
IV.
Discussion
Maternal susceptibility to preeclampsia is a multifactorial and complex process, involving the contribution of a
series of essential gene variants (Oudejans et al., 2007), affecting hemodynamic and vascular parameters. The
etiology and pathophysiology of preeclampsia are not fully understood, however oxidative stress has been
strongly linked to the occurrence of that disease (López-Jaramillo et al., 2001). The oxidative stress pathways
cause endothelial damage and induction of metabolic and coagulation aberrations. There is a suggested
involvement of glutathione S-transferases, major detoxificating enzymes, in the pathophysiology of
preeclampsia (Pappa et al., 2011). In the present study, we tried to assess a possible association between
glutathione S- transferase P1 -105 Ile →Val (-313 A to G) SNP (rs1695) as well as Mn-superoxide dismutase 16 Ala → Val (-47 C to T) SNP (rs4880) and patient risk for preeclampsia among Egyptian women.
DOI: 10.9790/3008-10132026
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Polymorphism In Glutatione S-Transferase P1 And Manganese-Superoxide Dismmutase…
The frequency of GSTP1-105 Ile/Val (AG) genotypes, Val/Val (GG) genotypes (both heterozygote and
homozygote forms ) and the frequency of carrying the Val (G) allele were significantly higher in preeclampsia
patients than control group. The relative risk of preeclampsia is higher in the carriers of Ile/Val (AG) and
Val/Val (GG) genotypes and the carriers of Val -105 (G) alleles than Ile-105 (A) allele. In accordance of the
observed results of the current study, some studies reported the same associations as the Val-105 (G) allele was
associated with an increased risk of developing preeclampsia in a Dutch population (Zusterzeel et al., 2000)
and with severe hypertension in elderly pregnancy in a Japanese population (Ohta et al., 2003). It is of interest
that homozygosity for the Val-105 polymorphism was associated with severe preeclampsia because of the
production of a GSTP1 enzyme with reduced detoxifying capacity (Zusterzeel et al., 2002).
However, a study performed by Canto et al. (2008) on Maya-Mestizo women reported different
associations in which the GSTP1-G (Val) allele conferred a reduced risk of preeclampsia under a dominant
model, since the comparison of AG and GG vs. AA genotype frequencies was significantly different. Also, no
association was found in the colored population of South Africa (Gebhardt et al., 2004). Moreover, Pappa et
al. (2011) speculated that the Val-105 allele frequency was higher in normal pregnancies than in preeclampsia
among Greek population. In fact, candidate gene studies suppose that single-gene effects may be variable across
populations (Zintzaras et al., 2006).
For Mn-superoxide dismutase-16 Ala → Val (-47 C to T) polymorphism, it was found that no
significant differences in the genotype frequencies and allelic distribution were detected among preeclampsia
and control groups (Table 4, 5). These findings are in accordance with the result of Zhang et al (2008) who
reported that the effects of the Ala-9Val are not fully understood and therefore it is difficult to hypothesize the
effects of the variant on the risk of preeclampsia. Also, Kim et al. (2005) found that there was no significant
association between Mn-SOD polymorphism and the risk for preeclampsia. Whereas, in contrary to our results,
Procopcius et al. (2012) reported significant association between Val/Val (TT) and preeclampsia in Romanian
population. No other studies were found for Mn-SOD polymorphism association to preeclampsia, so these
conflicting findings warrants further investigation in other populations.
In summary, we found that glutathione S- transferase P1 -105 Ile →Val (-313 A to G) SNP (rs1695) is
associated with the susceptibility to preeclampsia in pregnant Egyptian women. Whereas, our data did show any
significant difference between Mn-superoxide dismutase -16 Ala → Val (-47 C to T) SNP (rs4880) in the
samples taken from normal and preeclamptic pregnancies.
References
[1].
[2].
[3].
[4].
[5].
[6].
[7].
[8].
[9].
[10].
[11].
[12].
[13].
[14].
[15].
Aggarwal BB, Gehlot P (2009): Inflammation and cancer: how friendly is the relationship for cancer patients? Curr-OpinPharmacol., 9(4):351-69.
Bag A, Bag N (2008): Target sequence polymorphism of human manganese superoxide dismutase gene and its association with
cancer risk: a review. Cancer Epidemiol. Biomarkers Prev.,17(12):3298-305.
Cai Q, Shu XO, Wen W, Cheng JR, Dai Q, Gao YT, Zheng W (2004): Genetic polymorphism in the manganese superoxide
dismutase gene, antioxidant intake, and breast cancer risk: results from the Shanghai Breast Cancer Study. Breast Cancer Res., 6(6):
R647-55.
Canto P, Canto-Cetina T, Juárez-Velázquez R, Rosas-Vargas H, Rangel-Villalobos H, Canizales-Quinteros S, Velázquez-Wong
AC, Villarreal-Molina MT, Fernández G, Coral-Vázquez R (2008): Methylenetetrahydrofolate reductase C677T and glutathione Stransferase P1 A313G are associated with a reduced risk of preeclampsia in Maya-Mestizo women. Hypertens Res., 31(5):1015–19.
Chamy VM, Lepe J, Catalán A, Retamal D, Escobar JA, Madrid EM (2006): Oxidative stress is closely related to clinical severity of
pre-eclampsia. Biol Res., 39(2): 229–36.
Gebhardt GS, Peters WH, Hillermann R, Odendaal HJ, Carelse-Tofa K, Raijmakers MT, Steegers EA (2004): Maternal and fetal
single nucleotide polymorphisms in the epoxide hydrolase and gluthatione S−transferase P1 genes are not associated with preeclampsia in the Coloured population of the Western Cape, South Africa J. Obstet. Gynaecol., 24(8): 866–72.
Hurl SE, Lee JY, Moon HS, Chung HW (2005): Polymorphisms of the genes encoding theGSTM1, GSTT1 and GSTP1 in Korean
women: no association with endometriosis Mol. Hum. Reprod., 11 (1): 15-9.
Kariz S, Nikolajevic Starcevic J, Petrovic D (2012): Association of manganese superoxide dismutase and glutathione S-transferases
genotypes with myocardial infarction in patients with type 2 diabetes mellitus. Diabetes Res. Clin. Pract., 98(1): 144–50.
Kim YJ, Park HS, Park MH, Suh SH, Pang MG (2005): Oxidative stress-related gene polymorphism and the risk of preeclampsia.
Eur. J Obstet. Gynecol. Reprod. Biol., 119(1): 42-6.
Lain KY, Roberts JM (2002): Contemporary concepts of the pathogenesis and management of preeclampsia. JAMA; 287(24):
3183–6.
Li D, Dandara C, Parker MI (2010): The 341C/T polymorphism in the GSTP1 gene is associated with increased risk of
esophageal cancer. BMC Genetics, 11: 47-55.
López-Jaramillo P, Casas JP, Serrano N (2001): Preeclampsia: from epidemiological observations to molecular mechanisms. Braz.
J. Med. Biol. Res., 34(10): 1227–35.
Manica-Cattani MF, de Cadoná FC, Oliveira R, de Silva T, Machado AK, Barbisan F, Duarte MMF, da Cruz IBN (2012): Impact of
obesity and Ala16Val MnSOD polymorphism interaction on lipid, inflammatory and oxidative blood biomarkers. Open Genetics 2
(4): 202-9.
Ohta K, Kobashi G, Hata A, Yamada H, Minakami H, Fujimoto S, Kondo K, Tamashiro H (2003): Association between a variant
of the glutathione S−transferase P1 gene (GSTP1) and hypertension in pregnancy in Japanese: interaction with parity, age, and
genetic factors. Semin Thromb Hemost., 29(6): 653–59.
Oudejans CB, van Dijk M, Osterkamp M, Lachmeijer A, Blankenstein MA (2007): Genetics of preeclampsia: paradigm
shifts. Hum-Genet.,120(5): 607–12.
DOI: 10.9790/3008-10132026
www.iosrjournals.org
25 | Page
Polymorphism In Glutatione S-Transferase P1 And Manganese-Superoxide Dismmutase…
[16].
[17].
[18].
[19].
[20].
[21].
[22].
[23].
[24].
[25].
[26].
[27].
[28].
Pappa KI, Roubelakis M, Vlachos G, Marinopoulos S, Zissou A, Anagnou NP, Antsaklis A (2011): Variable effects of maternal
and paternal–fetal contribution to the risk for preeclampsia combining GSTP1, eNOS, and LPL gene polymorphisms. J- Matern
Fetal Neonatal Med., 24(4): 628–35.
Procopcyuc LM, Caracostea G, Nemeti G, Drugan C, Olteanu I, Stamatian F (2012): The Ala-9Val (Mn-SOD) and Arg213Gly (ECSOD) polymorphisms in the pathogenesis of preeclampsia in Romanian women: association with the severity and outcome
of preeclampsia. J. Matern.Fetal Neonatal. Med., 25(7):895-900.
Schur BC, Bjereke J, Nuwayhid N, Wong SH (2001): Genotyping of cytochrome P450-2D6*4 and *3 mutations using
conventional PCR.Clin.Chim. Acta, 308(1-2): 25 – 31.
Sutton A, Imbert A, Igoudjil A, Descatoire V, Cazanave S, Pessayre D, Degoul F (2005): The manganese superoxide dismutase Ala
16 Val dimorphism modulates both mitochondrial import and mRNA stability. Pharmacogenet-Genomics., 15(5): 311–9.
Tian C, Fang S, Du X, Jia C (2011): Association of the C47T polymorphism in SOD2 with diabetes mellitus and diabetic
microvascular complications: a meta-analysis. Diabetologia,54(4): 803–11.
Wang S, Wang F, Shi X, Dai J, Peng Y, Guo X, Wang X, Shen H, Hu Z (2009): Association between manganese superoxide
dismutase (MnSOD) Val-9Ala polymorphism and cancer risk-A meta-analysis. Eur J Cancer., 45(16): 2874-81.
Xu Z, Zhu H, Luk JM, Wu D, Gu D, Gong W, Tan Y, Zhou J, Tang J, Zhang Z, Wang M, Chen J (2012): Clinical Significance
of SOD2 and GSTP1 Gene Polymorphisms in Chinese Patients With Gastric Cancer . Cancer.,118(22): 5489-96.
Zelko IN, Mariani TJ and Folz RJ (2002): Superoxide dismutase multigene family: a comparison of the CuZn-SOD (SOD1), MnSOD (SOD2), and EC-SOD (SOD3) gene structures, evolution, and expression. Free Radic. Biol. Med., 33(3): 337-49.
Zhang J, Masciocchi M, Lewis D, Sun W, Liu A and Wang Y (2008): Placental anti-oxidant gene polymorphisms, enzyme activity,
and oxidative stress in preeclampsia. Placenta, 29(5): 439–43.
Zintzaras E, Kitsios G, Harrison GA, Laivuori H, Kivinen K, Kere J, Messinis I, Stefanidis I, Ioannidis JP (2006): Heterogeneitybased genome search meta-analysis for preeclampsia. Hum Genet.; 120(3):360–70.
Zusterzeel PL, Peters WH, De Bruyn MA, Knapen MF, Merkus HM, Steegers EA (1999): Glutathione S−transferase isoenzymes in
decidua and placenta of preeclamptic pregnancies. Obstet-Gynecol., 94(6): 1033–8.
Zusterzeel PL, Visser W, Peters WH, Merkus HW, Nelen WL, Steegers EA (2000): Polymorphism in the glutathione S−transferase
P1 gene and risk for preeclampsia. Obstet. Gynecol., 96(1): 50–4.
Zusterzeel PL, te Morsche R, Raijmakers MT, Roes EM, Peters WH, Steegers EA (2002): Paternal contribution to the risk for preeclampsia. J. Med. Genet., 39(1): 44–5.
DOI: 10.9790/3008-10132026
www.iosrjournals.org
26 | Page